Abstract

OBJECTIVESAlthough considerable progress has been made in regenerative medicine, a quantum step
would be the replacement and/or regeneration of functional muscle tissue. For example,
although patients' airways can now be successfully replaced with stem cell-based techniques,
a much greater patient need would be addressed by regeneration of the muscles required
for engineering a functional larynx, in which active movement is critical. The rabbit
cricoarytenoid dorsalis muscle was chosen for the present study because it is equivalent
to the posterior cricoarytenoid muscle, the only significant abductor muscle in human
larynges.METHODSRabbit cricoarytenoid dorsalis muscles were harvested, and different decellularization
methods were compared by use of a combination of histologic, immunohistochemical,
and molecular techniques. Decellularized scaffolds were implanted into Sprague-Dawley
rats as part of a 2-week biocompatibility study to assess immunogenicity.RESULTSDecellularization with a combination of latrunculin B, potassium iodide, potassium
chloride, and deoxyribonuclease resulted in total DNA clearance and reduced levels
of major histocompatibility complex class II expression, with relative preservation
of the scaffold's structural integrity (collagen, elastin, and glycosaminoglycan content).
The scaffolds showed minimal signs of rejection at 2 weeks in a cross-species (xenotransplantation)
study.CONCLUSIONSDecellularized laryngeal muscles, which are nonimmunogenic, may provide the optimal
scaffold source for the generation of a fully functional tissue-engineered larynx.